Preview - Magnolia Press

Zootaxa 3941 (3): 401–413 /zootaxa /
Copyright © 2015 Magnolia Press
ISSN 1175-5326 (print edition)
ISSN 1175-5334 (online edition)
A cultivable acoel species from the Mediterranean, Aphanostoma pisae sp. nov.
(Acoela, Acoelomorpha)
Research unit Evolutionary Developmental Biology, Institute of Zoology, University of Innsbruck, Technikerstr. 25, 6020 Innsbruck,
Austria. E-mail: [email protected], [email protected], [email protected]
Corresponding author. E-mail: [email protected]
Aphanostoma pisae sp. nov. is an interstitial acoel found at the coast of the Liguric Sea in Pisa (Tuscany, Italy). It belongs
to the large family Isodiametridae, characterised by a male copulatory organ with a cylindrical shape and non-anastomising longitudinal muscle fibers. It is the first recognised species of Aphanostoma in the Mediterranean and it can occur in
great abundance at its type locality (several hundred specimens in a spoonful of sand). A. pisae has been cultured in the
laboratory for several years with diatoms for food. The embryonic development lasts for just under two days at 20 °C.
We provide a description of the new species using live observations, light and electron microscopy of sagittal
sections and stainings of the filamentous actin and the serotonergic nervous system, and we discuss and update the genus
diagnoses of the genera Aphanostoma and Praeconvoluta.
Key words: Xenacoelomorpha, acoel flatworm, taxonomy, phalloidin, antibody stainings
Acoels are predominantly marine worms with a contested phylogenetic position. For more than a century, acoels
were considered members of the Platyhelminthes based on many morphological similarities (Ehlers, 1985), but a
great number of molecular analyses now place them either as sister group of the Bilateria (Ruiz-Trillo et al., 1999,
Egger et al., 2009) or, most recently, as deuterostomes (Philippe et al., 2011). Identification of acoels requires
careful observation and in many cases sagittal sections of the genital organs of mature animals are required for
identification (Westblad, 1948). Staining of the musculature (F-actin) can provide an additional level of detail for
characterising these organs and the body wall musculature (Hooge, 2001; Hooge & Tyler, 2005).
While many Scandinavian and Northern German acoel species have been described, the Mediterranean (and
even more so most other parts of the world) are still comparatively poorly covered (Nilsson et al., 2011). This trend
is reflected in the absence of recognised species of the genus Aphanostoma Ørsted, 1845 in the Mediterranean.
With this work, we provide the first description of an Aphanostoma species in the Mediterranean with images of
live animals, sagittal sections, and stainings of the musculature and the serotonergic nervous system. We also
propose a revision of the closely related genera Aphanostoma and Praeconvoluta Dörjes, 1968.
Material and methods
Sampling. Sand samples from Marina di Pisa, Italy (43.6761°N 10.2698° E) were taken in May 2005 and May
2011. Animals were extracted from the sand in 7% MgCl2 · 6H2O mixed 1:1 with artficial sea water and then
transferred to petri dishes (see below).
Cultures. Aphanostoma pisae was cultured in petri dishes with enriched sea water (f/2 medium) and fed ad
libitum with the diatom Nitzschia curvilineata in a constant environment at 20°C with a day/night cycle of 12/12
hours. About every two weeks, algae were replaced if necessary. When starting new dishes, about 40 adult worms
Accepted by W. Sterrer: 5 Mar. 2015; published: 1 Apr. 2015
Licensed under a Creative Commons Attribution License
Aphanostoma species (see Jondelius et al., 2011; Nilsson et al., 2011). Second, there is hardly any difference in the
generic diagnoses of Aphanostoma and Praeconvoluta, other than an elliptical penis sheath in Praeconvoluta, as
opposed to a spherical penis sheath in Aphanostoma. Molecular analyses so far suggest both genera should be
merged (see Jondelius et al., 2011; Nilsson et al., 2011). To help with this decision, it seems to be worthwhile to
make an effort constructing a phylogenetic tree of the Isodiametridae with as many species as possible.
In memoriam Reinhard Rieger, who told his students not to become taxonomists.
We are especially grateful to Matthew Hooge for his great help in species determination and reviewing the
manuscript and to two anonymous reviewers. We'd like to acknowledge all Pisa excursion participants in 2005 and
2011: Reinhard Rieger, Gunde Rieger, Robert Gschwentner, Johannes Achatz, Christian Gärber, Lukas Schärer,
Dita Vizoso, Gregor Schulte, Peter Sandner, Johanna Zaubzer, Lena Egger, Lena Zitzelsberger, Veronika Prantl,
Diane Rudulph and Julian Smith III; Johannes Achatz is also acknowledged for contributing valuable taxonomic
insights and for helping with the acknowledgements.
Achatz, J.G. & Martinez, P. (2012) The nervous system of Isodiametra pulchra (Acoela) with a discussion on the neuroanatomy
of the Xenacoelomorpha and its evolutionary implications. Frontiers in Zoology, 9, 27.
Beklemischev, V.N. (1915) Sur les turbellariés parasites de la côte Mourmanne. I. Acoela. Travaux de la Société impériale des
naturalistes de St. Petersbourg, 43 (4), 103–172.
Diesing, K.M. (1862) Revision der Turbellarien. Abtheilung: Rhabdocoelen. Sitzungsberichte der MathematischNaturwissenschaftlichen Classe der Kaiserlichen Akademie der Wissenschaften Abtheilung 1, 45, 191–318.
Dörjes, J. (1968) Die Acoela (Turbellaria) der deutschen Nordseeküste und ein neues System der Ordnung. Journal of
Zoological Systematics and Evolutionary Research, 6, 56–452.
Dörjes, J. (1972) Faerlea echinocardii sp. n. und Diskussion der Gattungen Avagina Leiper und Faerlea Westblad (Turbellaria
Acoela). Zoologica Scripta, 1 (3), 185–189.
Egger, B. & Ishida, S. (2005) Chromosome fission or duplication in Macrostomum lignano (Macrostomida,
Plathelminthes)—remarks on chromosome numbers in "archoophoran turbellarians". Journal of Zoological Systematics
and Evolutionary Research, 43 (2), 127–132.
Egger, B., Steinke, D., Tarui, H., De Mulder, K., Arendt, D., Borgonie, G., Funayama, N., Gschwentner, R., Hartenstein, V.,
Hobmayer, B., Hooge, M., Hrouda, M., Ishida, S., Kobayashi, C., Kuales, G., Nishimura, O., Pfister, D., Rieger, R.,
Salvenmoser, W., Smith III, J., Technau, U., Tyler, S., Agata, K., Salzburger, W. & Ladurner, P. (2009) To be or not to be a
flatworm: The acoel controversy. PLoS ONE, 4 (5), e5502.
Ehlers, U. (1985) Das Phylogenetische System der Plathelminthes. Gustav Fischer, Stuttgart & New York, 317 pp.
Eisenman, E.A. & Alfert, M. (1982) A new fixation procedure for preserving the ultrastructure of marine invertebrate tissues.
Journal of Microscopy, 125, 117–120.
Faubel, A. (1974) Die Acoela (Turbellaria) eines Sandstrandes der Nordseeinsel Sylt. Mikrofauna des Meeresbodens, 32, 1–58.
Faubel, A. (1977) Zwei neue interstitielle Arten der Acoela (Turbellaria) von Norwegen. Zoologica Scripta, 6 (1), 21–25.
Faubel, A. & Regier, S. (1983) Anatomy and taxonomy of new Acoela (Turbellaria) from the sublittoral of the northern North
Sea. Senckenbergiana maritima, 15 (1), 1–17.
Hooge, M.D. (2001) Evolution of body-wall musculature in the Platyhelminthes (Acoelomorpha, Catenulida, Rhabditophora).
Journal of Morphology, 249 (3), 171–194.
Hooge, M.D. & Rocha, C.E.F. (2006) Acoela (Acoelomorpha) from the northern beaches of the state of São Paulo, Brazil, and
a systematic revision of the family Otocelididae. Zootaxa, 1335, 1–50.
Hooge, M.D. & Tyler, S. (1999) Body-wall musculature of Praeconvoluta tornuva, n. sp. (Acoela, Platyhelminthes) and the use
of muscle patterns in taxonomy. Invertebrate Biology, 118 (1), 8–15.
412 · Zootaxa 3941 (3) © 2015 Magnolia Press
Hooge, M.D. & Tyler, S. (2003) Acoels (Platyhelminthes, Acoela) from the Atlantic coast of North America. Meiofauna
Marina, 12, 7–36.
Hooge, M.D. & Tyler, S. (2005) New tools for resolving phylogenies: a systematic revision of the Convolutidae
(Acoelomorpha, Acoela). Journal of Zoological Systematics and Evolutionary Research, 43 (2), 100–113.
Hooge, M.D. & Tyler, S. (2008) Acoela (Acoelomorpha) from Bocas del Toro, Panama. Zootaxa, 1719, 1–40.
Jensen, O.S. (1878) Turbellaria ad litora Norvegiae occidentalia. Turbellarier ved Norges Vestkyst. J.W. Eided Bogtrykkeri,
Bergen, 97 pp.
Jondelius, U., Wallberg, A., Hooge, M. & Raikova, O.I. (2011) How the worm got its pharynx: phylogeny, classification and
Bayesian assessment of character evolution in Acoela. Systematic Biology, 60 (6), 845–871.
Kozloff, E.N. (1965) New species of the acoel turbellarians from the Pacific coast. Biological Bulletin, 129, 151–166.
Ladurner, P. & Rieger, R.M. (2000) Embryonic muscle development of Convoluta pulchra (Turbellaria: Acoelomorpha,
Platyhelminthes). Developmental Biology, 222 (2), 359–375.
Meixner, J. (1938) Turbellaria (Strudelwürmer). In: Grimpe, G. & Wagler, E. (Eds.), Die Tierwelt der Nord- und Ostsee. Vol. 33
(IVb). Akademische Verlagsgesellschaft, Leipzig, pp. 1–146.
Mollenhauer, H.H. (1964) Plastic embedding mixtures for use in electron microscopy. Stain Technology, 39 (2), 111–114.
Nilsson, K.S., Wallberg, A. & Jondelius, U. (2011) New species of Acoela from the Mediterranean, the Red Sea, and the South
Pacific. Zootaxa, 2867, 1–31.
Nimeth, K.T., Egger, B., Rieger, R., Salvenmoser, W., Peter, R. & Gschwentner, R. (2007) Regeneration in Macrostomum
lignano (Platyhelminthes): cellular dynamics in the neoblast stem cell system. Cell and Tissue Research, 327, 637–646.
Ørsted, A.S. (1845) Fortegnelse over Dyr, samlede i Christianafjord ved Drøbak. Naturhistorisk Tidsskrift udgivet of Henrik
Krøyer, Series 2, 1, 400–427.
Pereyaslawzewa, S. (1892) Monographie des turbellaries de la Mer Noire. Schriften der neurussischen naturforschenden
Gesellschaft, 17 (3), 1–303.
Petrov, A., Hooge, M. & Tyler, S. (2006) Comparative morphology of the bursal nozzles in acoels (Acoela, Acoelomorpha).
Journal of Morphology, 267 (5), 634–648.
Philippe, H., Brinkmann, H., Copley, R.R., Moroz, L.L., Nakano, H., Poustka, A., Wallberg, A., Peterson, K.J. & Telford, M.J.
(2011) Acoelomorph flatworms are deuterostomes related to Xenoturbella. Nature, 470 (7333), 255–260.
Reuter, M., Raikova, O.I. & Gustafsson, M.K.S. (2001) Patterns in the nervous and muscle systems in lower flatworms.
Belgian Journal of Zoology, Supplement 1 (131), 47–53.
Richardson, K.C., Jarret, L. & Finke, E.H. (1960) Embedding in epoxy resins for ultrathin sectioning in electron microscopy.
Stain Technology, 35 (6), 313–323.
Romeis, B. (1968) Mikroskopische Technik. R. Oldenbourg Verlag, München, 757 pp.
Ruiz-Trillo, I., Riutort, M., Littlewood, D.T., Herniou, E.A. & Baguña, J. (1999) Acoel flatworms: earliest extant bilaterian
Metazoans, not members of Platyhelminthes. Science, 283 (5409), 1919–1923.
Salvenmoser, W., Egger, B., Achatz, J.G., Ladurner, P. & Hess, M.W. (2010) Electron microscopy of flatworms: standard and
cryo-preparation methods. Methods in Cell Biology, 96, 307–330.
Smith, J.P.S. & Bush, L. (1991) Convoluta pulchra, n. sp., (Turbellaria: Acoela) from the east coast of North America.
Transactions of the American Microscopical Society, 110, 12–26.
Spurr, A.R. (1969) A low-viscosity epoxy resin embedding medium for electron microscopy. Journal of Ultrastructure
Research, 26 (1–2), 31–43.
Steinböck, O. (1933) Die Turbellarienfauna der Umgebung von Rovigno. Thalassia, 1, 1–33.
Tyler, S. & Rieger, R.M. (1999) Functional morphology of musculature in the acoelomate worm, Convoluta pulchra
(Plathelminthes). Zoomorphology, 119 (3), 127–141.
Tyler, S., Schilling, S., Hooge, M. & Bush, L.F. (2006–2013) Turbellarian taxonomic database. Version 1.7. Available from: (accessed 1 November 2014).
Westblad, E. (1945) Studien über skandinavische Turbellaria Acoela. III. Arkiv för Zoologi, 36A (5), 1–56.
Westblad, E. (1948) Studien über skandinavische Turbellaria Acoela. V. Arkiv för Zoologi, 41A (7), 1–82.
Zootaxa 3941 (3) © 2015 Magnolia Press ·